A closed vitrification system enables a murine ovarian follicle bank for high-hroughput ovotoxicity screening, which identifies endocrine disrupting activity of microcystins

• Authors: Yingzheng Wang, University of South Carolina; Jingshan Xu, University of South Carolina; Jessica E Stanley, Oregon Natl Primate Res Ctr; Murong Xu, University of South Carolina; Bryan W Brooks, University of South Carolina; Geoffrey Scott, University of South Carolina; Saurabh Chatterjee, University of South Carolina; Qiang Zhang, Emory University; Mary B Zelinski, Oregon National Primate Research Center; Shuo Xiao, University of South Carolina
• Journal Title: REPRODUCTIVE TOXICOLOGY
• Volume: Volume 93
• Publisher: PERGAMON-ELSEVIER SCIENCE LTD | 2020-04-01, Pages 118-130
• Type of Work: Article | Post-print: After Peer Review
• Publisher DOI: http://doi.org/10.1016/j.reprotox.2020.01.009
• Subjects: Health Sciences, Public Health; Environmental Sciences
• Keywords: Science & Technology; Life Sciences & Biomedicine; Reproductive Biology; Toxicology; Vitrification; In vitro follicle growth; High-throughput; Ovotoxicity; Microcystin; IN-VITRO DEVELOPMENT; MOUSE PREANTRAL FOLLICLES; ZONA-PELLUCIDA GENES; PRE-ANTRAL FOLLICLES; PP2A INHIBITION; SECONDARY FOLLICLES; ALGINATE BEADS; HUMAN EXPOSURE; TISSUE; SPINDLE

Abstract

Increasing evidence reveals that a broad spectrum of environmental chemicals and pharmaceutical compounds cause female ovarian toxicity (ovotoxicity). The current gold standard of ovotoxicity testing largely relies on whole laboratory animals, but in vivo models are time consuming, costly, and present animal welfare concerns. We previously demonstrated that the 3D encapsulated in vitro follicle growth (eIVFG) is a robust in vitro model for ovotoxicity testing. However, the follicle preparation process is complex and highly dependent on technical skills. Here, we aimed to use vitrification method to cryopreserve murine immature follicles for a high-content eIVFG, chemical exposure, and ovotoxicity screening. Results indicated that a closed vitrification system combined with optimized vitrification protocols preserved mouse follicle viability and functionality and vitrified follicles exhibited comparable follicle and oocyte reproductive outcomes to freshly harvested follicles during eIVFG, including follicle survival and development, ovarian steroidogenesis, and oocyte maturation and ovulation. Moreover, vitrified follicles consistently responded to ovotoxic chemical, doxorubicin (DOX). We further used vitrified follicles to test the response of microcystins (MCs), an emerging category of environmental contaminants produced by cyanobacteria associated with harmful algal blooms (HABs), and found that different congeners of MCs exhibited differential ovotoxicities. In summary, our study demonstrates that vitrification enables a long-term-storage and ready-to-use ovarian follicle bank for high-throughput ovotoxicity screening, which identifies endocrine disrupting effects of MCs.